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Radiosensitive cell lines derived from X‐ray cross complementing group 5 (XRCC5), SCID mice and a human glioma cell line lack components of the DNA‐dependent protein kinase, DNA‐PK, suggesting that DNA‐PK plays an important role in DNA double‐strand break repair. Another enzyme implicated in DNA repair, poly(ADP‐ribose) polymerase, is cleaved and inactivated during apoptosis, suggesting that some DNA repair proteins may be selectively targeted for destruction during apoptosis. Here we demonstrate that DNA‐PKcs, the catalytic subunit of DNA‐PK, is preferentially degraded after the exposure of different cell types to a variety of agents known to cause apoptosis. However, Ku, the DNA‐binding component of the enzyme, remains intact. Degradation of DNA‐PKcs was accompanied by loss of DNA‐PK activity. One cell line resistant to etoposide‐induced apoptosis failed to show degradation of DNA …